DESCRIPTION

CD3 is a complex of proteins that directly associates with the T cell receptor (TCR). The TCR/CD3 complex of T-lymphocytes consists of either a TCR alpha/beta or TCR gamma/delta heterodimer coexpressed at the cell surface with the invariant subunits of CD3 labeled gamma, delta, epsilon, and zeta. The TCR recognizes antigens bound to major histocompatibility complex (MHC) molecules. TCR-mediated peptide-MHC recognition is transmitted to the CD3 complex, leading to the intracellular signal transduction. CD3E encodes the CD3-epsilon polypeptide which plays an essential role in T-cell development in addition to its role of signal transduction in T-cell activation. Defects in CD3E cause immunodeficiency.

PRINCIPLE OF THE ASSAY

A capture antibody is pre-coated onto the bottom of wells which binds to analyte of interest. A detection antibody also binds to the analyte. Horseradish peroxidase (HRP)-conjugated secondary antibody binds to the detection antibody. TMB acts as the HRP substrate and the solution color will change from colorless to blue. A stop solution containing sulfuric acid turns solution yellow. The color intensity is proportional to the quantity of bound protein which is measurable at 450 nm with the correction wavelength set at 630 nm.

PRECISION

Intra-assay Precision (Precision within an assay) Three samples of known concentration were tested 20 times on one plate to assess intra-assay precision. Inter-assay Precision (Precision between assays) Three samples of known concentration were tested in 24 separate assays to assess inter-assay precision.

Intra-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	20	2,083.4	163.6	7.9
2	20	454.2	31.2	6.9
3	20	119.1	6.9	5.8

Inter-assay Precision

Sample	n	Mean (pg/mL)	SD	CV%
1	24	1.825.7	162.8	8.9
2	24	452.6	26.1	5.8
3	24	115.6	11.3	9.8

RECOVERY

The recovery of human CD3E spiked to three different levels throughout the range of the assay in Cell lysate was evaluated.

Sample Type		Average% of Expected	Range (%)
Cell lysate	1:800	105	73-121
	1:1,600	104	87-121

LINEARITY

To assess the linearity of the assay, samples were diluted with the appropriate Sample Diluent to produce samples with values within the dynamic range of the assay. (The cell lysate was initially diluted 1:200)

		Cell lysate
1:2	Average% of Expected	100
	Range (%)	-
1:4	Average% of Expected	103
	Range (%)	93-115
1:8	Average% of Expected	119
	Range (%)	113-129
1:16	Average% of Expected	109
	Range (%)	96-124

CALIBRATION

This immunoassay is calibrated against a highly purified recombinant human CD3E produced at Sunlong Medical™.

Ordering & shipping

• Minimum order: 1 kit
• Lead time: varies with order quantity and specifications. Please consult your local distributor or our sales team for confirmation
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• Shipping: FedEx / DHL / UPS express; cold-chain packing available on request
• Price: USD, per kit (48T / 96T options available)